There is a need for new technologies to enable high-resolution, multiplexed analysis of protease activity. Advances in this area have the potential to enable the discovery of fundamental new insights into cellular function and regulation. We propose to develop a novel technology for sensitive and specific analysis of protease activity using protease substrate peptide microarrays. We will develop methods for array preparation, peptide labeling and detection that are optimized for high-throughput protease activity analysis. PUBLIC HEALTH RELEVANCE: The system we are developing for protease activity analysis will help to enable the deciphering of cellular regulation of protein degradation and the relationship to phenotype. It will help to identify disorders in cellular regulation that lead to disease, and therefore may result in medical advances in the diagnosis and treatment of disease.